LOCUS       sequence-270337-        5748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5748)
  AUTHORS   Stemmann O, Zou H, Gerber SA, Gygi SP, Kirschner MW
  TITLE     Dual inhibition of sister chromatid separation at metaphase.
  JOURNAL   Cell. 2001 Dec 14;107(6):715-26.
  PUBMED    11747808
REFERENCE   2  (bases 1 to 5748)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell."; 
            date: "2001-12-14"; volume: "107(6)"; pages: "715-26"
COMMENT     Sequence Label: pCS2-hSecurin
FEATURES             Location/Qualifiers
     source          1..5748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1087..1260
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1261..1434
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1454..1465
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1477..1497
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1513..1533
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1585..1605
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1621..1641
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2754..2773)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2755..2773)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2778..2912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2802..2821)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2856..2875
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3025..3045)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3025..3043)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3064..3080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3064..3080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3077..3099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3088..3104
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3112..3118,3119..3136,3137..3142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3157..3178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3295..3312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3466..4054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3546..3565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4225..5016,5017..5085))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4848..4867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5086..5190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5216..5671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5353..5374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5565..5584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccgccatggc gcaacacgat
     1081 gaagccgtag acaacaaatt caacaaagaa caacaaaacg cgttctatga gatcttacat
     1141 ttacctaact taaacgaaga acaacgaaac gccttcatcc aaagtttaaa agatgaccca
     1201 agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgctca ggcgccgaaa
     1261 gtagacaaca aattcaacaa agaacaacaa aacgcgttct atgagatctt acatttacct
     1321 aacttaaacg aagaacaacg aaacgccttc atccaaagtt taaaagatga cccaagccaa
     1381 agcgctaacc ttttagcaga agctaaaaag ctaaatgatg ctcaggcgcc gaaagtagac
     1441 gcgaattcgc tcgatcgagg gccgcccggc agcagcgaga acctgtactt ccagggcgcc
     1501 atccccacca ccgagaacct gtacttccag agcggcaccg tggatgccgg cgccgatgcc
     1561 ggcaagcccc ggcccggcag cagcgagaac ctgtacttcc agggcgccat ccccaccacc
     1621 gagaacctgt acttccagag cggcaccgtg gatgccggcg ccgatgccgg caagccccgg
     1681 ccgatgagga gcttcaaaag aggccggccg atggctactc tgatctatgt tgataaggaa
     1741 aatggagaac caggcacccg tgtggttgct aaggatgggc tgaagctggg gtctggacct
     1801 tcaatcaaag ccttagatgg gagatctcaa gtttcaacac cacgttttgg caaaacgttc
     1861 gatgccccac cagccttacc taaagctact agaaaggctt tgggaactgt caacagagct
     1921 acagaaaagt ctgtaaagac caagggaccc ctcaaacaaa aacagccaag cttttctgcc
     1981 aaaaagatga ctgagaagac tgttaaagca aaaagctctg ttcctgcctc agatgatgcc
     2041 tatccagaaa tagaaaaatt ctttcccttc aatcctctag actttgagag ttttgacctg
     2101 cctgaagagc accagattgc gcacctcccc ttgagtggag tgcctctcat gatccttgac
     2161 gaggagagag agcttgaaaa gctgtttcag ctgggccccc cttcacctgt gaagatgccc
     2221 tctccaccat gggaatccaa tctgttgcag tctccttcaa gcattctgtc gaccctggat
     2281 gttgaattgc cacctgtttg ctgtgacata gatatttaag gcgcgccatc tagaaatgaa
     2341 ggtgtcgcaa cttatgcagc tgctgttctg ttccgtatgt ctgaggacaa accccaggac
     2401 tacaagaagc gtctgtcagt tgagttgacc agctctctct tcaggaccga gccaatgcca
     2461 tggaatgagg ctgcagacct tggtcttgat attggtgccc aaggcgaacc tcttggctac
     2521 agacaggatg attcaagcta ccgatctttc catgctgctg gctatggtca ggatgcaatg
     2581 ggtatggact ccatgatgga tcatgacatg ggaggtcacc acccaggggc agactatcca
     2641 gttgatggac ttcctgattt gagtcatgcc caagatctta tggatggact acctccaggt
     2701 gatagcaacc aactggcctg gtttgacact gacttgtaaa ctagattctg cagccctata
     2761 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     2821 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2881 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2941 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     3001 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3061 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3121 agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5221 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5281 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5341 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5401 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5461 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5521 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5581 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5641 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5701 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgacc
//
